alpha-Mannosidosis Mutation Database
A database on mutations, genotypes and the clinical and molecular aspects of alpha-Mannosidosis

Experimental Result

Western blot analysis, transfected COS-7 cells

Cells were transfected with the appropriate MAN2B1-construct or empty vector and harvested 36-48 h post-transfection. Ten μg of protein (lysate) or 10 μl of medium was analysed by SDS/PAGE (4–12% gel) under reducing conditions and electroblotted onto a PVDF membrane. MAN2B1 was detected using antiserum against native recombinant human MAN2B1 and ALP-conjugated secondary antibody followed by chemiluminescence detection.

( Riise Stensland et al. 2012 )

Schematic view on the MAN2B1 sequence, proteolytic processing and structure.

Xtra Image

The three dimensional structure contains 5 domains, the alpha/beta barrel, 3alpha-bundle and three beta-domains beta1-beta3. The disulphide bridges and the 11 potential glycosylation sites on human MAN2B1 are marked on the structure panel. The proteolytic sites are modelled from the bovine MAN2B1 structure (1o7d, ref). In addition to the peptides a-d, the structure contains two additional proteolytic cuts, which create small peptides d’ and e’.